Hosotsubo K K, Takezawa J, Nishijima M K, Takahashi H, Shimada Y, Yoshiya I
Crit Care Med. 1985 Dec;13(12):1061-3. doi: 10.1097/00003246-198512000-00016.
Varying concentrations of lipopolysaccharide (LPS) and mannan suspensions were mixed with either saline or plasma from normal volunteers, heated to 100 degrees C for 10 min, and then subjected to the limulus amebocyte lysate test (LAL). A positive LAL in saline required minimum LPS and mannan concentrations of 10(-11) and 10(-5) g/ml, respectively, while the minimum concentrations premixed with plasma were 10(-13) and 10(-9) g/ml, respectively. Thus, use of plasma instead of saline increased assay sensitivity 100-fold for LPS and 10,000-fold for mannan. In the second part of the experiment, normal plasma was separated into lipid and nonlipid phases by Folch's method. LAL analysis of each phase revealed equal sensitivity for LPS and mannan in the nonlipid phase, but no sensitivity in the lipid extract. Subsequently, 200 ml of a fat emulsion (Intralipos) was administered to the normal volunteers, and LAL and lipid analyses were performed. The LAL turned positive in all volunteers. When LAL was positive, triglycerides (TG), chylomicron (Chyl), and very low-density lipoprotein (VLDL) increased significantly compared with when LAL was negative. It is concluded that plasma lipids increase the sensitivity of LAL and directly activate LAL when TG, Chyl, and VLDL concentrations are high. This effect of plasma lipids on LAL can be eliminated by extracting LPS and mannan in the nonlipid phase.
将不同浓度的脂多糖(LPS)和甘露聚糖悬浮液与正常志愿者的生理盐水或血浆混合,加热至100摄氏度10分钟,然后进行鲎试剂检测(LAL)。生理盐水中LAL呈阳性时,所需的最低LPS和甘露聚糖浓度分别为10^(-11)和10^(-5)克/毫升,而与血浆预混合时的最低浓度分别为10^(-13)和10^(-9)克/毫升。因此,用血浆代替生理盐水可使LPS检测灵敏度提高100倍,使甘露聚糖检测灵敏度提高10000倍。在实验的第二部分,通过Folch法将正常血浆分离为脂质相和非脂质相。对各相进行LAL分析发现,非脂质相中LPS和甘露聚糖的灵敏度相同,但脂质提取物中无灵敏度。随后,给正常志愿者静脉输注200毫升脂肪乳剂(英脱利匹特),并进行LAL和脂质分析。所有志愿者的LAL均呈阳性。LAL呈阳性时,甘油三酯(TG)、乳糜微粒(Chyl)和极低密度脂蛋白(VLDL)与LAL呈阴性时相比显著升高。得出的结论是,当TG、Chyl和VLDL浓度较高时,血浆脂质会增加LAL的灵敏度并直接激活LAL。血浆脂质对LAL的这种影响可通过在非脂质相中提取LPS和甘露聚糖来消除。